Placental bed biopsies taken during caesarean section from 10 patients with pre-eclampsia and six healthy pregnancies were studied. We applied antibodies against cytokeratin and different macrophage markers to analyse the distribution of invasive extravillous trophoblast cells as compared to that of macrophages in myometrial segments of uteroplacental arteries. The data were evaluated quantitatively. We found a clear inverse relationship between local infiltration with macrophages and trophoblast invasion. In pre-eclampsia, vessel cross-sections prevailed which were characterized by large numbers of macrophages but a low degree of trophoblast invasion. In contrast, in normal third trimester pregnancies the respective arterial segments had a high degree of trophoblast invasion but were largely void of macrophages. These data suggest causal links between macrophages and inhibition of intra-arterial trophoblast invasion in pre-eclampsia.

Adult↗

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Phage display: a molecular tool for the generation of antibodies--a review.

Phage display comprises an array of methods, which can be used to display proteins on bacteriophages. We present in this article a summary of techniques, which can be used to express antibody libraries on bacteriophages. Since many researchers are more familiar with the conventional hybridoma technique for production of monoclonal antibodies we describe analogies and differences between these two techniques, both of which are used to reach comparable scientific objectives. We focus on the features which are specific to phage display techniques rather than for hybridoma techniques. These comprise the freedom to choose other animals than the mouse for immunization, the enormously large sample (up to 10(9) clones) which can be drawn and immortalized from a single immunized animal and the opportunity to enhance affinity of isolated antibodies by in vitro affinity maturation. The panning techniques, which are used to enrich specifically binding phage antibodies from the huge libraries are briefly summarized.

Animals↗

PubMedDate not supplied

Cell culture models of human trophoblast--primary culture of trophoblast--a workshop report.

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Cell Culture Techniques↗

PubMedDate not supplied

Characterization of trophoblast cell isolations by a modified flow cytometry assay.

The purity of isolated trophoblast cells is an important checkpoint in in vitro studies on the human placenta. Maintaining viability of primary cells for a prolonged period, or achieving cell proliferation in primary cell cultures, is often a matter of concern. In this paper we present a method for characterizing the purity of isolated trophoblast cells based on the expression of cytoskeletal proteins, as well as for assessing their cell cycle status, by flow cytometry. We show that after a simple permeabilization and fixation step in 70 per cent methanol, staining for cytokeratin 7 and vimentin could discriminate between trophoblast cells and contaminating populations. The method was applicable to trophoblast cells both from villous and extravillous origin. By staining the proliferation-related antigen Ki-67 and DNA, information was gained about the cell cycle status and viability of freshly isolated and cultured villous trophoblast cells. This method may help to quickly and quantitatively characterize preparations of isolated trophoblast, as well as to search for culture conditions favouring long-term survival and proliferation.

Cell Cycle↗